LOCUS       sequence-353221-        6372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MTFR1L-V5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6372)
  AUTHORS   Schaffer BE, Levin RS, Hertz NT, Maures TJ, Schoof ML, Hollstein PE,
            Benayoun BA, Banko MR, Shaw RJ, Shokat KM, Brunet A
  TITLE     Identification of AMPK Phosphorylation Sites Reveals a Network of 
            Proteins Involved in Cell Invasion and Facilitates Large-Scale 
            Substrate Prediction.
  JOURNAL   Cell Metab. 2015 Nov 3;22(5):907-21. doi: 
            10.1016/j.cmet.2015.09.009. Epub 2015 Oct 8.
  PUBMED    26456332
REFERENCE   2  (bases 1 to 6372)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cmet.2015.09.009"; journalName: "Cell Metab"; date: 
            "2015-11-3- 3"; volume: "22"; issue: "5"; pages: "907-21"
COMMENT     Sequence Label: pcDNA 3.2/V5-DEST MTFR1L-V5
FEATURES             Location/Qualifiers
     source          1..6372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        627..830
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     875..894
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        875..893
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    923..947
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1824..1848)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1874..1915
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1955..1974)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1999..2047
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2249..2677
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2336..2355)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2546..2567
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2686..2706)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2691..3020
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2871..3006
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2933..2952
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3087..3881
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3141..3160)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3751..3770
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4057..4178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4094..4113)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4148..4167
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4227..4243)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4227..4243)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4240..4262)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4251..4267
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4275..4281,4282..4299,4300..4305))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4320..4341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4458..4475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4629..5217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4709..4728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5388..6179,6180..6248))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6011..6030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6249..6353)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      841 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      901 aagctggcta gttaagctat caacaagttt gtacaaaaaa gttggcatgt caggaatgga
      961 agccactgtg accatcccaa tctggcaaaa caagccacat ggggctgctc gaagtgtagt
     1021 aagaagaatt gggaccaacc tacccttgaa gccgtgtgcc cgggcgtcct ttgagaccct
     1081 gcccaacatc tctgacctgt gtttgagaga tgtgccccca gtccctaccc tggctgacat
     1141 cgcctggatt gctgcggatg aagaggagac atatgcccgg gtcaggagtg atacgcgccc
     1201 cctgaggcac acctggaaac ccagccctct gattgtcatg cagcgcaatg cctctgttcc
     1261 caacctgcgt gggtccgagg agaggcttct ggccctgaag aagccagctc tgccagccct
     1321 aagccgcact actgagctgc aggacgagct gagccacttg cgcagccaga ttgcaaagat
     1381 agtggcagct gatgcagctt cggcttcatt aacgccagat ttcttatctc caggaagttc
     1441 aaatgtctct tctcccttac cttgttttgg atcctcattc cactctacaa cttcctttgt
     1501 cattagtgac atcaccgagg agacagaggt ggaggtccct gagcttccat cagtccccct
     1561 gctttgttct gccagccctg aatgttgcaa accagaacac aaagctgcct gcagttcgtc
     1621 tgaagaggat gactgcgtct ctttgtccaa ggccagcagc tttgcagaca tgatgggtat
     1681 cctgaaggac tttcaccgaa tgaaacagag tcaagatctg aaccggagtt tattgaagga
     1741 ggaagaccct gctgtgctta tctctgaggt cctaaggagg aagtttgctc taaaggaaga
     1801 agatatcagt agaaaaggaa attgcccaac tttcttgtac aaagtggttg atctagaggg
     1861 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     1921 cggttagtaa tgagtttaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     1981 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     2041 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     2101 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     2161 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     2221 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2281 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2341 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2401 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2461 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2521 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2581 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2641 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2701 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2761 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2821 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2881 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2941 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3001 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3061 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3121 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3181 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3241 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3301 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3361 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3421 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3481 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3541 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3601 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3661 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3721 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3781 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3841 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     3901 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     3961 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4021 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4081 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4141 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4201 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4261 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4321 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4381 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4441 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4501 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4561 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4621 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4681 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4741 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4801 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4861 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4921 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4981 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5041 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5101 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5161 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5221 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5281 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5341 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5401 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5461 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5521 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5581 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5641 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5701 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5761 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5821 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5881 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5941 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6001 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6061 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6121 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6181 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6241 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6361 ccccgaaaag tg
//
